KR101481778B1 - Composition containing the extract of ginseng berry containing increased ginsenoside Re for improving sexual function - Google Patents

Abstract

A composition containing an extract of a ginseng berry including increased ginsenoside Re for improving sexual function according to the present invention increases generation of nitrogen monoxide (NO) in vascular endothelial cells and shows a strong relaxing action in a smooth muscle of the corpus cavernosum to have an advantage of effectively alleviating male sexual disorders like erectile dysfunction. The composition of the present invention can be used as a pharmaceutical composition or a food composition for improving male sexual function.

Description

TECHNICAL FIELD The present invention relates to a composition for improving sexual function comprising ginsenoside Re-enriched ginseng fruit extract,

The present invention relates to a composition for improving sexual function comprising ginseng fruit extract, and more particularly, to a composition for improving sexual function, which comprises extracting ginsenoside Re-enhanced extract from ginseng fruit using ultrafiltration will be.

The ginseng was named after the Soviet scientist CAMeyer in 1843 as " Panax ginseng CA Meyer "is a botanical plant belonging to the genus Araliaceae (Panax), and its roots are used medicinally. Such ginseng is widely used as a natural health food, and has scientific proof of pharmacological efficacy and clinical , The recognition and trust is high, and the demand for medicine and functional food is increasing.

The ginseng saponin, which is a ginseng saponin, is mainly involved in ginseng saponins. The present ginsenoside has about 30 chemical structures. The main ginsenosides currently known are 13 basic ginsenosides and converted ginsenosides. Thirty-seven ginsenosides are known in eleven types of senosides. These ginsenosides exhibit various pharmacological effects. Depending on the chemical structure, the ginsenosides can be classified into a protopanaxadiol (PPD) such as ginsenosides Rb1, Rb2, Rc and Rd and a trioside such as ginsenosides Re, Rg1 and Rf , And protopanaxatriol (PPT).

Ginsenoside Re is a PPT family of ginsenoside that has been shown to have various effects such as central nervous system inhibitory action, anti-diabetic activity, corticosteroid stimulating action, and anti-stress action.

Ginseng consists of root, head, stem, leaf, and fruit. Ginseng fruit is harvested around July - August, and peel off flesh to obtain ginseng seed for sowing. Flesh of ginseng fruit is soft and has a characteristic of retreating if left as it is. Generally, fresh ginseng fruit is put into a peeler, and the ginseng flesh is broken by frictional force and is washed with water to collect seeds. Most of the ginseng flesh that is generated during the process of harvesting seeds is used for abandonment or for farming.

In addition, since it is diluted with water during the peeling process, it contains a considerable amount of water in comparison with the dry weight. Therefore, when it is concentrated by the conventional evaporation concentration, the ginsenoside Re, which is a main component of the ginseng fruit, And is destroyed by the organic acid contained therein. It is not easy to concentrate diluted ginseng fruit pulp, which is tethered with water, because ginsenoside Re has a characteristic that sugar chains are easily broken off by organic acids and lose the characteristics of saponin.

Further, when the ginseng pulp is concentrated by the conventional evaporation concentration method, it has a sticky liquid phase, which makes it difficult to form tablets and granules.

Erectile dysfunction is a phenomenon in which the penis does not persist even if the penis is not sufficiently erected, resulting in more than 25% of the total sex life, and normal blood flow into the corpus cavernosum can not be achieved (Goldstein et al., 1998). The cause is classified into cardiogenic and metachronous, and substrate is classified into neurogenic, endocrine, vascular, and systemic diseases (Goldstein et al., 1998). Neurogenic erectile dysfunction is caused by brain tumors, cerebrovascular disease, spinal cord injury, peripheral neuropathy caused by diabetes or chronic alcohol poisoning (Benet et al., 1995).

Korean Patent No. 10-0857767 entitled " Method for Producing Ginseng Extract from Ginseng Fruit ", the ginseng fruit treated with light is mixed with steam and dried, and then the extract is prepared by using hot water extraction method, organic solvent extraction method and supercritical extraction method Method is described. However, this method has a problem that the ginsenoside Re present in the fruit of ginseng is decomposed by heat and the ginsenoside content in the final extract is lowered.

Korean Patent No. 10-1030608 discloses a composition for improving male sexual function, which is prepared by extracting ginseng fruit with water or alcohol in "composition for improving male sexual function containing ginseng fruit extract". However, since the above composition is merely a method for producing an extract from ginseng fruit using hot water extraction method or alcohol extraction method, a specific ginsenoside which can prevent, treat and improve male sexual dysfunction such as erectile dysfunction contained in ginseng fruit There is no description about a method capable of selectively strengthening the components.

KR 10-1030608 B1, 14 April 2011

Accordingly, the inventors of the present invention discovered a process for producing ginsenoside Re-enriched ginseng fruit extract while continuing the study to solve the above problems, and found that a ginsenoside Re-enhanced ginseng fruit extract containing enriched ginseng fruit extract The present invention has been completed by developing a composition.

The object of the present invention is to provide a composition for improving sexual function using a process for extracting and concentrating ginsenoside Re, which is a major component of ginseng fruit, as well as a process for producing ginseng fruit extract having enhanced powdery ginsenoside Re . The present invention provides a composition for improving sexual function comprising ginsenoside Re-enriched ginseng fruit extract, which is characterized by a method for producing a ginseng fruit extract having enhanced ginsenoside Re using ultrafiltration.

Another object of the present invention is to provide a composition for improving sexual function comprising ginsenoside Re-enriched ginseng fruit extract.

Yet another object of the present invention is to provide a composition for improving sexual function comprising ginsenoside Re-enriched ginseng fruit extract, which is characterized by using the composition as a pharmaceutical composition or food composition for the prevention and treatment of male erectile dysfunction .

To achieve the object of the present invention, there is provided a composition for improving sexual function comprising ginsenoside Re-enriched ginseng fruit extract.

In order to achieve the above object, the present invention provides a composition for improving sexual function comprising ginsenoside Re-enriched ginseng fruit extract, which comprises dissolving a flesh of ginseng fruit to prepare ginseng flesh fluid; Dissolving the ginseng fruit pulp component of the ginseng flesh liquid and centrifuging it; Filtering the centrifuged supernatant; And ultrafiltering said filtrate to a molecular weight of 1000-2000 to obtain a concentrate.

The composition for improving sexual function comprising ginsenoside Re-enriched ginsenoside extract of the present invention provides a ginsenoside Re-enriched ginseng fruit extract used as a pharmaceutical composition or food composition for preventing and treating male erectile dysfunction .

Hereinafter, the present invention will be described in detail.

In the present invention, after completion of the step of obtaining the concentrate of the above-mentioned production method, the concentrate may further be dried by a high-efficiency centrifugal concentrator or freeze drier and pulverized. In one embodiment of the present invention, it may be dried at 80 to 100 ° C for 2 to 6 hours.

In the present invention, the ginseng pulp liquid preparation step can be manufactured by putting the ginseng fruit into a desorber and adding water to the pulp, and removing the pulp from the seed.

In this case, the ginseng fruit may be subjected to a pretreatment such as drying or steaming. However, in one embodiment of the present invention, fresh ginseng fruit is used, and the ginseng fruit is put into a degasifier. The ginseng powder is crushed by frictional force, Flesh liquid can be produced

In the present invention, in the centrifuging step, the ginseng pulp may be left as it is or ultrasonically for 30 minutes to 2 hours, preferably 1 hour to elute the components of the ginseng fruit pulp and then centrifuge.

In this case, the centrifugal separator is usually used in the art and is not limited to the kind.

In the present invention, in the filtration step, the supernatant may be obtained by centrifugal separation, and then filtered on a filter paper by a conventional method.

In the present invention, in the step of obtaining the concentrated liquid, the filtrate is passed through an ultrafiltration membrane having a molecular weight of 1,000 to 2,000, preferably a molecular weight of 1000 to obtain a concentrate (inner liquid) of 15 to 30% It can be concentrated to about 20%.

In the present invention, the ginseng fruit extract may be concentrated using an reverse osmotic pressure concentrator (nanofilter system) instead of using an ultrafiltration membrane in the step of obtaining the concentrate. In this case, Side Re. ≪ / RTI >

The production method of the present invention can improve the content of ginsenoside Re by about 1% to 10% as compared with the conventional concentration method.

In addition, the production method of the present invention may show a ginsenoside content of the PPT series about 2 to 5 times higher than the conventional concentration method (for example, evaporation vacuum concentration method, freeze drying method, etc.).

In the present invention, in the centrifuging step, the ginseng pulp liquid is centrifuged at 15,000 rpm in a continuous high-speed centrifuge, and in the step of obtaining the concentrated liquid, freezing-drying the concentrated liquid obtained by the ultrafiltration is further included can do.

The composition for improving sexual function comprising the ginsenoside Re-enriched ginsenoside extract of the present invention may be formulated as a pharmaceutical composition or a food composition in the form of tablets, pills, tablets, capsules, granules, powders, ointments, It can be changed.

The composition may be prepared into a pharmaceutical formulation according to a conventional method. In the preparation of the formulations, it is preferred that the active ingredient is mixed with or diluted with the carrier, or enclosed in a carrier in the form of a container. When the carrier is used as a diluent, it may be a solid, semi-solid or liquid substance acting as a carrier, excipient or medium for the active ingredient. Thus, the formulations may be in the form of tablets, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, sterile injections, sterile powders and the like. The compositions of the present invention may be formulated using methods well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal.

Herein, the pharmaceutical composition of the present invention can be administered through various routes including oral, transdermal, subcutaneous, intravenous, intraperitoneal, muscle, topical application, patch and iontophoresis, . The actual dosage of the active ingredient of the present invention should be determined in view of various factors such as the disease to be treated, the route of administration, the age, sex and weight of the patient, and the severity of the disease, But is not limited to.

When the composition of the present invention is prepared as a food composition, it contains not only an extract of ginseng fruit as an active ingredient but also a component which is ordinarily added during the manufacture of a food, for example, a protein, a carbohydrate, a fat, a nutrient, . Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, .

The composition for improving sexual function comprising ginsenoside Re-enriched ginsenoside extract according to the present invention increases nitric oxide (NO) production in vascular endothelial cells and exhibits a strong relaxation action in the smooth muscle of cavernosal corpus cavernosum, There is an advantage of effectively improving sexual dysfunction.

FIG. 1 shows an HPLC chromatogram of a ginsenoside Re-enriched ginseng fruit extract according to the present invention.
FIG. 2 shows the effects of the ginsenoside Re-enriched ginseng fruit extract according to the present invention on body weight, dietary fiber and insulin content of the erectile dysfunction model (A: weekly body weight change, B: weekly dietary intake, C: ).
FIG. 3 shows the effect of the ginsenoside Re-enriched ginseng fruit extract according to the present invention on diurnal blood glucose changes in the erectile dysfunction model.
4 is a graph showing an analysis of the internal pressure of the penile corpus cavernosum.
Figure 5 shows the effect of ginsenosides Re is enhanced ginseng fruit extract on the corpus cavernosum pressure of ED model according to the invention ^{(a, b, c, d} ) a statistical significance level of the test group by the mean value is p < 0.05 for each subgroup (n = 6)).
Figure 6 shows the effect of ginsenosides Re is enhanced ginseng fruit extract on the corpus cavernosum NO content of ED model according to the invention ^{(a, b, c, d} ) a statistical significance level of the test group by the mean value of p <0.05 for each subset (n = 6)).
Figure 7 shows the effect of ginsenosides Re is enhanced ginseng fruit extract on the corpus cavernosum cGMP content of ED model according to the invention ^{(a, b, c, d} ) a statistical significance level of the test group by the mean value of p <0.05 for each subset (n = 6)).

The present invention will be described in more detail with reference to the following examples. However, the following examples are provided to aid understanding of the present invention, and the scope of the present invention is not limited by these examples in any sense.

Hereinafter, the technical and scientific terms used herein will be understood by those skilled in the art without departing from the scope of the present invention. Descriptions of known functions and configurations that may be unnecessarily blurred are omitted.

In the embodiment of the present invention, the ultrafiltration was performed using a Quixstand benchtop system (GE Healthcare, UK) and a filter of 1,000 NMWC hollow fiber cartridge (GE Healthcare, UK Model: UFP-1-C-4M) Respectively.

In the examples of the present invention, Nano filter system (Youngin Co., Korea) was used for the reverse osmosis concentration and Dow Filmtec (USA) and NF30-1812-24 (44.5 x 298H) Respectively.

Analysis of the ginsenoside content of each extract was carried out by using water (liquid A) and ACN (liquid B) as a solvent using a Waters-PDA (Waters 1525, detector 2998, USA) The same analytical conditions were analyzed by high performance liquid chromatography (HPLC) at a flow rate of 1.0 ml / min.

Time
(min) Flow (ml / min) H2O (%)
(Liquid A) ACN
(Liquid B) 1.0 82.0 18.0 42 1.0 76.0 24.0 46 1.0 71.0 29.0 79 1.0 60.0 40.0 115 1.0 35.0 65.0 135 1.0 15.0 85.0 150 1.0 15.0 85.0 151 1.0 82.0 18.0

Control factor Condition HPLC Waters-PDA (Waters 1525, detector 2998) Injection volume 20 μL Column Xbridge ™ C18
(250x4.6 mm 5 [mu] m, Waters) Mobile phase Solvent A: Water, Solvent B: ACN Flow rate 1.00 mL / min Column Temperature 40 ℃ Wavelength 203 nm

[ Experimental Example  One] Gin Senocide Re  Comparative analysis of production method of Ginseng fruit extract

1. Ethanol precipitation method Gin Senocide Re  Manufacture of ginseng fruit extract

20 kg of ginseng fruit collected from Geumsan was put into a fruit harvesting machine, and then the fruit was sieved. The remaining flesh was freeze-dried to obtain pulp powder. Powdered pulp powder was weighed 10 times 70% ethanol and extracted 3 times at room temperature for 3 days. After the extract was filtered, the filtrate was concentrated under reduced pressure until the filtrate became 20%, then left to stand in a refrigerator for one night. The precipitate to be precipitated was placed on a filter paper and filtrated under reduced pressure to freeze-dry the filtercake to obtain ginsenoside Re fraction .

As shown in Table 3 below, the weight of the freeze-dried powder remaining after removing the seeds from 20 kg of ginseng fruit was 1.5 kg, and a yield of about 7.5% was obtained. The purity of Re was 5.0% or more. The extract weight of 70% ethanol was 1.3 kg, which was 6.5% of the yield of ginseng. The fraction of ginsenoside Re obtained from the ethanol extract was about 50 g, and the yield of ginseng was 0.25% Re purity was more than 20%.

2. In ultrafiltration  Following Gin Senocide Re  Manufacture of ginseng fruit extract

20 kg of ginseng fruit was put into a peeler, water was added, and the pulp was removed from the seed to obtain 50 L of ginseng pulp. Ginseng powder is left as it is or ultrasonically for 1 hour to dissolve the components of ginseng fruit and then centrifuge to obtain supernatant, which is then filtered through a filter paper. The filtrate was passed through an ultrafiltration membrane having a pore size of 1000 molecular weight to concentrate the concentrate (internal solution) which did not pass through the ultrafiltration membrane to 20%. This was dried using a high-efficiency centrifugal concentrator (Genevac, UK) or a freeze dryer (Ilshin, Korea) to obtain 500 g of the ginseng fruit extract powder (B). Ultrafiltration equipment was a Quixstand benchtop system (GE Healthcare, UK) equipped with a hollow fiber cartridge (Model: UFP-1-C-4M, type: 1,000 NMWC, GE Healthcare, UK).

3. Heating of ginseng fruit extract

20 kg of ginseng fruit was put into a peeler, water was added, and the pulp was removed from the seed to obtain 50 L of ginseng pulp. The ginseng pulp was left as it was or ultrasonically for 1 hour to elute the components of ginseng fruit pulp and centrifuged to obtain supernatant, which was then filtered through a filter paper. 5 L of 50 L of the filtrate was lyophilized (A). To confirm the change of the components of ginseng fruit, it was left at 90 ° C for 3 hours and then lyophilized to obtain 150 g of each ginseng fruit extract.

4. Manufacture of ginseng fruit extract by conventional concentration method

20 kg of ginseng fruit was put into a peeler, water was added, and the pulp was removed from the seed to obtain 50 L of ginseng pulp. The ginseng pulp was left as it was or ultrasonically for 1 hour to elute the components of ginseng fruit pulp and centrifuged to obtain supernatant, which was then filtered through a filter paper. 5 L of 50 L of the filtrate was concentrated by evaporation (60 ° C) in an evaporator as the conventional method to obtain 150 g of ginseng fruit extract.

5. Depending on the concentration method Gin Senocide  Content change

The contents of ginsenosides according to the freeze-drying, ultrafiltration, and evaporation vacuum concentration methods of ginseng fruit pulp water extracts were compared (Table 4). The ginsenoside Re content of the ginseng fruit extract was 50.14 mg / g, which was about 5%, by using the freeze-dried concentrate which was not subjected to any heat at all. The ginseng fruit extract which did not pass through the ultrafiltration membrane prepared using the ultrafiltration membrane The ginsenoside Re content of the concentrate (inner solution) was 64.55 mg / g, about 6.5%, and the ginsenoside Re content of the ginseng fruit extract by the conventional evaporation vacuum concentration method was about 4.2%, which was 41.55 mg / g . Extraction method by ultrafiltration was able to produce ginsenoside Re - fortified ginseng fruit extract with a content of ginsenoside Re of about 1.5% higher than freeze - drying and 2.3% higher than the conventional evaporation vacuum concentration method. In addition, PT (protopanaxatriol) ginsenoside was found to be 3 to 4 times higher than that of PT. When preparing a concentrate by reverse osmosis concentrator (nanofilter system) Similar ginsenoside Re was contained.

6. Heating of Ginseng Fruit Extract Gin Senocide  Content change

The ginseng fruit extract contained a large amount of organic acid and was recorded at pH 4.5. Organic acid is used as a biocatalyst to break the 20 sugar chain of ginseng saponin and change ginsenosides Rb1, Rb2, Rc and the like as PD (protopanaxadiol) saponins to Rg3, Rk1, Rg5 and the like. PT (protopanaxatriol) The sugar chain is cut off and the non-saponin chain PT is made through Rh1 and the like. Therefore, when the ginseng fruit extract containing a large amount of organic acid is heated, the ginsenoside Re, which is a major component of the ginseng fruit, is rapidly decomposed. The ginsenoside Re content was found to decrease by 60% from 50.14 mg / g before heating to 20.02 mg / g after heating at 90 ℃ for 3 hours and then lyophilization. (Table 5). Therefore, it is possible that the concentration of ginsenoside Re, which is an effective ingredient of ginseng fruit extract, is very low and the content of ginsenoside Re in the final concentrate is very low or very low. From this viewpoint, the method for producing ginsenoside Re-fortified ginsenoside Re by ultrafiltration or nanofilter system is a method for producing ginseng fruit extract having physiological activity using ginsenoside Re as an active ingredient.

7. Comparison of extraction yields of extracts from ginseng fruit juice

To compare the yields of ginseng fruit pulp extracted from ginseng fruit according to the extraction solvent and extraction method, ① ginseng fruit juice was lyophilized, or ② 1x distilled water or ③50% ethanol (EtOH) was added to the fruit juice, The centrifuged supernatant was centrifuged at reduced pressure, or ④ 1 × water was added to the ginseng fruit juice, and the supernatant obtained by high-speed centrifugation at 15,000 rpm was dried to obtain the yield from the fruit juice.

The yields of ginseng fruit juice after removing seeds from ginseng fruit were 5.46% when lyophilized ginseng fruit juice was lyophilized, and 1 times of distilled water was added to fruit juice and centrifuged at 3000 rpm When the supernatant was concentrated under reduced pressure, 1.78%, ③50% ethanol (EtOH) was added and the concentration of the centrifugal supernatant was reduced to 2.08% at 3000 rpm (Table 6). The extraction yield of 50% ethanol was higher than that of water extraction. ④ The yields obtained by concentrating the supernatant obtained by high speed centrifugation at 15,000 rpm by adding 1 time of water to the ginseng fruit juice were much higher than 1.78% when centrifuged at a low rate of 2.37%.

8. After continuous centrifugation, extracts of ginseng fruit Gin Senocide Re  Stability test of the components against heat

To examine the stability of ginseng fruit juice, 200ml of ginseng fruit juice was centrifuged (15000rpm) and vacuum dried (evapration, 65 ℃) and the other was lyophilized as nectar. The other one was boiling in a 100 ° C water bath for 30 minutes and lyophilized to determine the dry weight of each and analyze the ginsenoside Re content.

As shown in Table 7, the ginsenoside Re content was the highest when lyophilized, and decreased by 22% when boiling. When it was dried under reduced pressure, it was reduced by about 4% compared with the case of freeze drying. After centrifugation, the drying yield of the supernatant was about 2.35%.

9. Continuous centrifuged fruit Supernatant  Of powder Thermal stability  Review

The thermal stability of ginsenoside Re was examined by measuring the content of ginsenoside Re by heating in a dry oven (105 ° C, 1 hr) freeze-dried or evaporated powder of the supernatant of continuous centrifugation.

As shown in Table 8, there was no significant difference in the ginsenoside Re content between the ginseng powder powder dried at reduced pressure and the freeze-dried powder at 105 ° C for 1 hour before and after heating.

10. Ginseng fruit juice powder (30% dextrin  Addition) manufacturing

For the commercialization of ginseng fruit juice powder, ginseng fruit juice was continuously centrifuged, concentrated at 50% decompression, and spray - dried with 30% dextrin to prepare ginseng fruit juice powder.

As shown in Table 9, when the content of ginsenoside in the powder was 30% dextrin added to the powder by spray drying and general vacuum drying, the content of ginsenoside of 30% dextrin- Re content decreased from 9.6% to 4.6% by about 50%.

[ Experimental Example  2] Gin Senocide  Content analysis

[ Example  One] In ultrafiltration  Following Gin Senocide Re  Manufacture of ginseng fruit extract

After centrifugation (15,000 rpm) in a continuous high-speed centrifuge, 1 L of the supernatant was concentrated to 50% with a 1 kDa pore size ultrafiltration membrane (220 RPM, 15 psi) was lyophilized and powdered to obtain 75 g of a sample. Hereinafter, a sample for evaluation of sexual function is used and is referred to as 'GS-F3K1'.

[ Comparative Example  1] Production of red ginseng extract

1 kg of 70% ethanol was added to 1 kg of 4: 6 mixture of 4-year-old red ginseng root and red ginseng purchased from Geumsan Ginseng Nonghyup and extracted three times at 70 ° C for 6 hours, followed by drying under reduced pressure to obtain 135 g of a sample . Hereinafter, referred to as 'REG'.

One. In ultrafiltration  Following Gin Senocide Re  Ginseng fruit extract extract Gin Senocide  Component analysis

As shown in Table 10 below, the ginsenoside Re content before and after the ultrafiltration was 8.6% before the ultrafiltration and increased to 10.9% after the ultrafiltration by about 26%.

2. GS -F3K1 and REG of Gin Senocide  Comparison of content analysis

As shown in the following Table 11, the ginsenoside content of GS-F3K1 was ginsenoside Rg1 7.0, Re 109.0, Rb1 13.2, Rc 10.5, Rb2 12.4, Rd 12.1 mg / g. The ginsenosides content of red ginseng concentrate (REG) used for positive control was ginsenoside Rg1 7.4, Re 12.4, Rf 3.9, Rb1 32.6, Rc 22.7, Rb2 9.8, Rd 4.6 mg / g. It was confirmed that the content of ginsenoside Re of the ginseng fruit extract (GS-F3K1) according to the present invention was enhanced.

[ Experimental Example  3] Experimental effect on sexual function in alcohol-induced erectile dysfunction model

Based on the above results, the effect of improving the male reproductive function of the ginseng fruit extract according to the present invention was tested. Specific experimental methods are as follows.

One. Test group  Isolation and administration

The rats undergoing the refinement period were weighed, separated evenly according to the Z - array method, and identified by using ear punch. The ginseng fruit fraction (0.25 g / kg) and GS-F3K1 (0.25 g / kg) were administered to the test group in the normal, control and low dose groups (ginseng fruit fraction 0.1 g / kg and GS-F3K1 0.1 g) (0.5 g / kg, GS-F3K1 0.5 g) and positive control (red ginseng extract 0.5 g / kg, RGE 0.5 g) were orally administered during the experimental period. Control and experimental groups were fed with 20% alcohol mixed water and each sample during the administration period. Twelve animals were tested in each group.

2. Test method and statistical processing

In the erectile dysfunction model using 6-week-old Sprague-Dawley (SD) rats, each test group was divided into two groups: normal, control, low dose group (ginseng fruit fraction 0.1 g / kg, GS-F3K1 0.1 g) (Ginseng fruit fraction, 0.5 g / kg, GS-F3K1 0.25 g), high concentration group (ginseng fruit fraction 0.5 g / kg and GS-F3K1 0.5 g), positive control (red ginseng extract 0.5 g / kg, RGE 0.5 g) Respectively. Control and experimental groups were fed with 20% alcohol mixed water and each sample during the administration period. After 5 weeks of oral administration, each test substance was analyzed for blood components by blood sampling, and the maximum intracapone pressure, NO and cGMP contents were measured.

Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by post-test with Ducan's multiple range test at p <0.05.

3. Measure weight, diet and food quantity

Body weights and weekly blood sugar changes were measured at a certain time a week. Dietary and insulin levels were measured after the feeding.

As shown in FIG. 2, the body weight, the dietary intake and the amount of water in each experimental group were lower in the alcohol group than in the normal group, but the difference between the control group and the ginseng fruit fraction- Was not confirmed.

4. Analysis of blood components

Serum samples collected from the abdominal vein at the time of animal autopsy were analyzed for serum albumin, aspartate transaminase (ALT), alanine transaminase (ALT), total cholesterol (TC), triglyceride (TG), low density lipoprotein-cholesterol ), High density lipoprotein-cholesterol (HDL-C), and γ-glutamyltransferase (γ-GTP) were analyzed and analyzed by the Green Cross Medical Foundation (GC Labs, Korea).

As shown in Table 12 below, there was no significant difference in the hematological analysis of erectile dysfunction model by drinking alcohol compared to the control group. In addition, as shown in FIG. 3, no significant difference in blood glucose level between the experimental groups was found.

5. Penetrating Cavernosal Pressure Measurement

Anesthesia was induced by intraperitoneal injection of xylazine 23.3 mg / ml (rumpun, Bayer animal drug) and ketamine 57.68 mg / ml (ketamine, Yuhan) at a ratio of 0.4 ml / Phenobarbital (Ruminal, Daihan Pharm Co., Ltd., Seoul, Korea), known as the sedative with the lowest blood pressure, was administered intramuscularly to maintain anesthesia. Heparinzed saline (100 IU / ml) was filled in Poly Ethylen 50 (Dow Corning, Midland, MO, USA) to prevent blood clotting and the silicone tube was connected to the carotid artery using a pressure transducer (MLT1199BP Transducer, AD Instruments, Colorado Springs, CO, USA). The pelvic ganglion and the cavernous cavernosal nerve were dissected using the same method as that of the cavernosal neurosurgery.

Platinum electrodes were placed on the cavernosal nerve for nerve stimulation and connected to Stimulator HC (ML155, AD Instrument). In addition, the penis perforation was incised to expose the corpus cavernosum, and a 22 G injection needle was placed in one corpus cavernosum to measure the cavernosal pressure. The pressure measurement needle was connected to a channel 2 of a pressure transducer (MLT1199BP Transducer, AD Instrument) and the electrical stimulation was controlled with a differential amplifier (Podbridge, AD Instrument) and Data Acquisition system (ML846 Powerlab 4/26, AD Instrument) And measurements were recorded and analyzed using a Data Analysis program (Powerlab Program, AD Instruments, USA).

The penile foot function test was performed by applying perforating nerve stimulation (frequency: 5 Hz, intensity: 3-5V, pulse duration: 500 pulses) for 1 minute to determine the erection standard for each animal. After that, the internal pressure of the cavernosal dropped to the basal level, and 10 minutes later, electric stimulation of the same intensity was applied and the paw function was measured three times, and the mean maximum cavernosal pressure was measured. Maximal corpus cavernosal pressure was expressed as maximal cavernosal pressure / mean arterial blood pressure.

Among the many factors that cause erectile dysfunction, alcohol decreases the ability of the erectile dysfunction, and irreversible prostate atrophy in alcoholic men leads to atrophy of the vas deferens and loss of sperm cells (Kurt, 1997; Gheng et al., 2007) . In order for the penis to eradicate, complete relaxation of the penis cavernosal artery and penile smooth muscle and accumulation of blood in the sinusoidal space within the corpus cavernosum (Chchiris, 1995) increases the corpus cavernosal pressure.

As shown in FIGS. 4 and 5, the maximal penile sponge pressure was observed after 5 weeks of ginseng fruit fraction administration to observe the erectile performance of the penis, and the maximum penile sponge pressure was corrected using mean arterial blood pressure. The blood pressure of the corpus cavernosum in the control group (control: 20% alcohol only) was 0.58 ± 0.02 mmHg, which was significantly lower than that of the normal group (0.76 ± 0.03 mmHg) (p <0.05). On the other hand, the intracellular Ca ^ <2 +> intrinsic pressure of the ginseng fruit fraction was significantly increased. Especially, in the case of GS-F3K1 0.5 g in the high concentration of ginseng fraction, 0.75 ± 0.01 mmHg in the corpus cavernosum pressure was used as a positive control ( P <0.05) than that of 0.66 ± 0.02 mmHg of red ginseng extract (RGE 0.5 g). Therefore, it was observed that the highest concentration of ginseng fruit fraction in the experimental group which received 20% alcohol had the highest intracoronary pressure, which is similar to that of the other experimental groups.

In this experiment, the change in the maximal penile sponge pressure was measured by forced oral administration of the ginseng fruit extract according to the present invention while drinking the alcohol for 5 weeks in the negative water. From the above results, it was confirmed that the decreased intracoronary pressure of corpuscle according to the administration of alcohol was restored in a concentration dependent manner upon administration of the ginseng fruit extract.

6. NO ( Nitrite , NO ₂ ^- ) Content analysis

NO has been reported to play a role in blood pressure control, neurotransmission, blood coagulation, and immune function as well as in vascular endothelial cells, as well as in neuron terminals and smooth muscle (Stamler et al., 1992) CGMP, an inactive substance generated from guanosine triphosphate by activated and activated guanyl cyclase, induces smooth muscle relaxation (Ignarro et al., 1990). To date, NO has been shown to be a major signaling agent for penile erection, confirming that the injection of nonspecific NOS inhibitors into the penile corpuscle of rats inhibits electrical nerve-induced penile erection in a dose-dependent manner, , And is known to be regulated by NOS (Knispel et al., 1992). The synthesized NO diffuses into smooth muscle cells and converts GTP to cGMP by guanylyl cyclase. Activated cGMP reduces intracellular calcium concentration and relaxes and expands cavernosal smooth muscle and small arteries to maintain erection (De et al. 1981).

The amount of nitrite (NO ₂ ^? ) In the tissues was analyzed by the manufacturer's recommended method using the colorimetric nitric oxide detection kit (INTRON Biotechnology, Inc., Gyeonggi, Korea) based on the Griess reaction. After adding 100 μl of the tissue supernatant to 96 wells, 50 μl of sulfanilamide solution (N1 buffer) was added and reacted at room temperature for 5 minutes. Then, 50 μl of Naphthylenediamine solution (N2 buffer) was added to each well, and after 5 minutes, it was measured at a wavelength of 540 nm using a spectrophotometer.

As shown in FIG. 6, the contents of NO (Nitrite, NO ₂ ^- ) in the corpus cavernosum were 9.9 ± 0.1 μmole in the control group and 11.4 ± 0.1 μmole in the control group. On the other hand, the NO content of the experimental group treated with the ginseng fruit fraction was significantly increased compared to the control group, which was 10.4 ± 0.1 μmole in the GS-F3K1 0.1 g test group, 11.0 ± 0.0 μmole in the GS-F3K1 0.25 g test group and 0.1 g in the GS-F3K1 test group (Fig. 4, p < 0.05). In particular, the GS-F3K1 0.5 g test group showed higher levels of NO than the normal and positive control groups. Based on the fact that NO was found to be the main neurotransmitter in penile erection, It is thought to be effective in inducing erection by expanding blood vessels.

Therefore, the content of erectile dysfunction related factors in the corpus cavernosum of the ginseng fruit extract group in the erectile dysfunction model was analyzed, and it was confirmed that the change of NO and cGMP contents by alcohol drinking was effectively improved.

7. cGMP  Content analysis

cGMP kit (Enzo Life sciences, Inc., FL, USA). 100 μl of tissue supernatant was added to a 96-well plate of cGMP kit, and 50 μl of conjugate and 50 μl of anti-body were added and incubated at 500 rpm for 2 hours at room temperature. After washing 3 times with washing buffer, 200 μl of pNpp substrate solution was added and reacted at room temperature for 1 hour. After stopping the reaction by adding 50 μl of stop solution, the absorbance was measured at 405 nm using a spectrophotometer.

As shown in FIG. 7, in the analysis of cGMP content in erectile dysfunction model, 1.8 ± 0.1 pmole of normal group was significantly higher than 0.9 ± 0.1 pmole of control group, indicating that cGMP level in corpus cavernosum tissue was decreased by drinking alcohol . These changes were improved by 1.2 ± 0.0 pmole of GS-F3K1, 1.7 ± 0.1 pmole of GS-F3K1, and 1.4 ± 0.0 pmole of GS-F3K1 and 0.5 g of GS-F3K1 and GS-F3K1 fractions, respectively Compared with the control group, the content of cGMP in corpus cavernosum was significantly increased ( p <0.05).

From the above results, it can be seen that the ginsenoside Re-enriched extract according to the present invention significantly increases the intracellular pressure of the corpuscles decreased by alcohol and increases the content of NO and cGMP in the corpus cavernosum, To improve erectile dysfunction.

Claims (4)

Removing ginseng fruit from the fruit to produce ginseng pulp;
The ginseng flesh fluid is left to stand for 30 minutes to 2 hours on an ultrasonic wave to elute the components of the flesh of the ginseng fruit and then centrifuged at 15,000 rpm in a continuous high speed centrifuge;
Filtering the centrifuged supernatant;
Ultrafiltering the filtrate to a molecular weight of 1000 to obtain a concentrate having a concentration of the concentrate (internal solution) of 15 to 30% which has not passed through the ultrafiltration membrane; And
The present invention relates to a composition for improving male sexual function comprising a ginsenoside Re-enriched ginseng fruit extract, which comprises the step of freeze-drying the concentrate obtained by the ultrafiltration. delete The method according to claim 1,
Wherein the composition is a pharmaceutical composition for the prevention and treatment of male erectile dysfunction, comprising a ginsenoside Re-enriched ginseng fruit extract. The method according to claim 1,
A composition for improving male sexual function comprising a ginsenoside Re-enriched ginseng fruit extract, wherein the composition is a food composition for preventing and improving male erectile dysfunction.

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